MicroRNAs are minor constituents of extracellular vesicles that are rarely delivered to target cells.
MicroRNAs are minor constituents of extracellular vesicles that are rarely delivered to target cells.
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DOI：
10.1371/journal.pgen.1009951
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发表时间：
2021-12
期刊：
影响因子：
4.5
通讯作者：
Hammerschmidt W
中科院分区：
文献类型：
--
作者：
Albanese M;Chen YA;Hüls C;Gärtner K;Tagawa T;Mejias-Perez E;Keppler OT;Göbel C;Zeidler R;Shein M;Schütz AK;Hammerschmidt W
哺乳动物细胞释放不同类型的囊泡，统称为细胞外囊泡（EV）。EV含有细胞微小RNA（miRNA），其具有将其miRNA货物递送至受体细胞以影响单个mRNA和细胞转录组的稳定性的明显潜力。miRNA通过EV途径输出的程度以及它们是否有助于细胞间通讯是有争议的。为了解决这些问题，我们定义了EV的多种特性，并分析了它们将包装的miRNA递送到靶细胞中以发挥生物学功能的能力。我们应用定义明确的方法来产生和表征具有或不具有特异性病毒miRNA的纯化EV。我们发现只有一小部分EV携带miRNA。EV很容易与不同的靶细胞类型结合，但EV不能与细胞膜可检测地融合以递送其货物。我们将EV设计成融合的，并记录了它们递送功能性信使RNA的能力。然而，工程融合EV没有可检测地改变暴露于携带miRNA的EV的细胞的功能。这些结果表明，EV携带的miRNA不作为细胞间通讯的效应子。大多数后生动物细胞释放不同类型和来源的囊泡，如外泌体和微囊泡，现在统称为细胞外囊泡（EV）。EV已经获得了很多关注，因为它们含有microRNA（miRNAs），因此可以在接受EV的受体或受体细胞中调节它们的特异性mRNA靶标。使用一种具有上级灵敏度和特异性的新型融合试验，我们重新审视了这一说法，但没有发现EV在许多不同细胞系和原代人血细胞中有效功能性摄取的令人信服的证据。即使是被设计成融合并将其miRNA货物递送到受体细胞的EV，在非常仔细控制的定量实验中也对靶mRNA没有可测量的影响。我们的阴性结果清楚地表明，EV并不能作为基于miRNA的细胞间通讯的载体。
Mammalian cells release different types of vesicles, collectively termed extracellular vesicles (EVs). EVs contain cellular microRNAs (miRNAs) with an apparent potential to deliver their miRNA cargo to recipient cells to affect the stability of individual mRNAs and the cells’ transcriptome. The extent to which miRNAs are exported via the EV route and whether they contribute to cell-cell communication are controversial. To address these issues, we defined multiple properties of EVs and analyzed their capacity to deliver packaged miRNAs into target cells to exert biological functions. We applied well-defined approaches to produce and characterize purified EVs with or without specific viral miRNAs. We found that only a small fraction of EVs carried miRNAs. EVs readily bound to different target cell types, but EVs did not fuse detectably with cellular membranes to deliver their cargo. We engineered EVs to be fusogenic and document their capacity to deliver functional messenger RNAs. Engineered fusogenic EVs, however, did not detectably alter the functionality of cells exposed to miRNA-carrying EVs. These results suggest that EV-borne miRNAs do not act as effectors of cell-to-cell communication. The majority of metazoan cells release vesicles of different types and origins, such as exosomes and microvesicles, now collectively termed extracellular vesicles (EVs). EVs have gained much attention because they contain microRNAs (miRNAs) and thus could regulate their specific mRNA targets in recipient or acceptor cells that take up EVs. Using a novel fusion assay with superior sensitivity and specificity, we revisited this claim but found no convincing evidence for an efficient functional uptake of EVs in many different cell lines and primary human blood cells. Even EVs engineered to fuse and deliver their miRNA cargo to recipient cells had no measurable effect on target mRNAs in very carefully controlled, quantitative experiments. Our negative results clearly indicate that EVs do not act as vehicles for miRNA-based cell-to-cell communication.
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DOI：
10.3390/s16070950
发表时间：
2016-06-23
期刊：
Sensors (Basel, Switzerland)
影响因子：
--
作者：
Jones DM;Padilla-Parra S
通讯作者：
Padilla-Parra S
DOI：
10.1126/science.aau6977
发表时间：
2020-02-07
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Kalluri R;LeBleu VS
通讯作者：
LeBleu VS
影响因子：
16.2
作者：
CORY, AH;OWEN, TC;CORY, JG
通讯作者：
CORY, JG
影响因子：
21.3
作者：
Hergenreider, Eduard;Heydt, Susanne;Dimmeler, Stefanie
通讯作者：
Dimmeler, Stefanie
影响因子：
7.3
作者：
Andreu Z;Yáñez-Mó M
通讯作者：
Yáñez-Mó M